Phytochemical Characterisation and LC-MS Profiling of Ziziphus mauritiana Leaf Extracts for Anti-inflammatory Potential
Background: Inflammation is the immune system’s first reaction to an injury or infection. Corticosteroids, immunosuppressants, and Non-Steroidal Anti-Inflammatory Drugs (NSAID) often used to treat inflammatory illnesses have serious adverse effects, such as cardiovascular, renal, and gastrointestinal damage. To lessen the negative effects of currently available medications, there is an urgent need to create innovative anti-inflammatory regimens derived from natural sources. Aim: The current study evaluated Ziziphus mauritiana for its anti-inflammatory potential, as it contains numerous bioactive constituents like phenolics, flavonoids, and saponins, which can contribute to the development of safer and cost-effective plant-based therapeutics for the treatment of inflammation. Methods: Methanol, hexane and ethyl acetate leaf extracts were evaluated for anti-inflammatory potential using LPS-stimulated RAW 264.7 macrophages at concentrations of 3-100 µg/mL for 24 hours. Nitric oxide production was evaluated by the Griess assay. Liquid Chromatography- Mass Spectrometry (LC-MS) analysis for characterisation of various phytoconstituents was determined on a Waters Alliance e2695 High Performance Liquid Chromatography (HPLC) system using a gradient of water, acetonitrile, methanol, and acetic acid. Results: Qualitative estimation shows the presence of phenols, triterpenoids, flavonoids and cardiac glycosides in methanol and ethyl acetate extract. In-vitro cytotoxicity evaluation demonstrated that the methanol and ethyl acetate extracts maintained more than 50% cell viability at 3 to 20 µg/mL, whereas the hexane extract showed reduced viability, indicating toxicity. Nitric oxide production in RAW 264.7 macrophages was inhibited significantly by methanol and ethyl acetate extract, with maximum inhibition at a concentration of 20µg/mL. LC-MS profiling of ethyl acetate fraction has shown the presence of various bioactive compounds such as L-carnitine, Delphinidin, caffeine, ononin, saccharopine, sinapine, Kaempferol-7-O-neohesperidoside, fortunellin, diosmin, marein etc. Conclusion: Results indicate that leaves of Z. mauritiana are rich in bioactive compounds and exhibit significant anti-inflammatory potential, mediated by quenching nitric oxide production, highlighting their potential for the development of plant-derived anti-inflammatory medications that are safe to use and have minimal side effects. Major Findings: Ziziphus mauritiana leaf extract has shown significant anti-inflammatory potential by the inhibition of nitric oxide production in the RAW 264.7 cell line. The anti-inflammatory potential of this plant may be attributed to the presence of various bioactive compounds such as L-carnitine, Delphinidin, caffeine, sinapine, Kaempferol-7-O-neohesperidoside, fortunellin, diosmin, marein, etc., analysed by LC-MS.